الصفحة 1
الصفحة 1
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Neutrophil : Methods and Protocols

Neutrophils, the most abundant white cells in humans, serve as the primary cellular defense against infection. This volume provides a concise set of protocols for assessing basic neutrophil functions and investigating specialized areas in neutrophil biology. Each of the protocols is written by leading researchers in the field and includes hints for success, as well as guidance for troubleshooting. Part I contains overviews of neutrophil biology, function, and disorders. Part II describes commonly used methods to isolate neutrophils from humans and other animal species. Part III details methods for investigating chemotaxis, transmigration, phagocytosis, and bactericidal activity. Part IV includes protocols that measure neutrophil apoptosis, calcium signal transduction, degranulation and detection of cytoplasmic granules, gene expression, transcription factors, and apoptosis. Part V provides multiple assays for measuring production of intracellular and/or extracellular reactive oxygen species, and a method that details use of the cell-free NADPH oxidase assay, an iconic assay for studies of the neutrophil NADPH oxidase. Part VI provides chapters that describe how to analyze formation and function of neutrophil extracellular traps.

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Neuroprotection : Method and protocols

Contains cutting-edge molecular biology methods on neuroprotective mechanisms and specific preclinical models of the CNS injury, iseases and planning translation. Chapters guide readers through neuropathology, neuroprotection, Alzheimer’s disease, amyotrophic lateral sclerosis, ALS, Huntington’s disease , multiple sclerosis, Parkinson’s disease, spinal cord injury, traumatic brain injury, and ischemic brain injury. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

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Neuronal cell death : Methods and protocols

Covers comprehensive methods on ways to assess structural and ultrastructural changes in the mitochondria, cytoskeleton, and microglia using state-of-the-art microscopy techniques including super-resolution imaging, electron microscopy, and ultra-high field MRI. The chapters in this book cover topics such as analysis of neurodegeneration in the post-mortem characterization of preclinical animal models, in vivo modeling in cell death in different model systems and brain organoids, single cell clonal analysis using Mosaic Analysis with Double Markers in genetic mouse models, and genome and proteomic methods for analysis of mRNA dynamics and quantitation of targeted peptides.

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Natural product experiments in drug discovery

Explores a wide range of evidence-based complementary medicine and various bio-analytical techniques used to define botanical products. Collecting recent work and current developments in the field of contemporary phytomedicine as well as their future possibilities in human health care, the book includes unique contributions in the form of chapters on phytomedicine and screening biological activities explained with diverse hyphenated techniques, as well as issues related to herbal medications, such as efficacy, adulteration, safety, toxicity, regulations, and drug delivery. Written for the Springer Protocols Handbooks series, chapters feature advice from experts on how to best conduct future experiments. Extensive and practical, Natural Product Experiments in Drug Discovery serves as an ideal reference for students, professors, and researchers in universities, R&D institutes, pharmaceutical and herbal enterprises, and health organizations.

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Mucins : Methods and protocols

Explores the latest advancements in mucin research. The chapters in this book are organized into 8 parts and cover a wide range of topics such as mucin extraction, isolation, physicochemical property analysis, and experimental methods. The chapters also discuss the origins of mucins in jellyfish, feces, saliva and salivary glands, bronchi, stomach, and cervical tract; organic synthesis of peptides glycosylated at specific sites; analysis of mucin gene expression and methylation-specific electrophoresis of genes; imaging of mucin networks by AFM; and experimental methods using supported molecular matrix electrophoresis. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

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Micropropagation methods in temporary immersion systems

Presents vital techniques for the commercial micropropagation of plants. It explores numerous micropropagation protocols in different types of temporary immersion systems for plant species with agricultural, medicinal, ornamental, and forestry interest, detailing the type of bioreactor and the optimal parameters necessary to guarantee obtaining the largest number of commercial propagules. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

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Microfluidics diagnostics : Methods and protocols

Explores a wide range of microfluidic-based approaches that exploit the unique features of microfluidic devices, which hold significant potential in the field of diagnosis. Beginning with a section on microchips for sample preparation, including cell-free DNA, exosomes, and cells, the book continues by covering protein marker analysis and detection, single-cell analysis, analysis of bacteria and viruses, as well as human cell-based culture and analysis. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

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Memory B-cells : Methods and protocols

Details in methods to assess memory B cell formation and function in mice and humans. Chapters guide readers through tetramer-based methods to assess antigen-specific memory B cell dynamics in humans and mice in different vaccine, next-generation deep-sequencing, single-cell techniques to assess epigenomic, VDJ landscapes, lymph node aspirates from humans, advanced imaging, murine models to determine memory B cells formation, and bioinformatic techniques and in silico modelling of memory b cell formation. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.

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Iron metabolism : Methods and protocols

Explores classical and cutting-edge methods optimized and validated to analyze various aspects of iron metabolism, from in vitro to multi-organ level. Opening with a section on basic iron metabolism methods, the book continues with methods applicable to a variety of systems, ranging from bacteria to cultured mammalian cells and tissues, with a focus on cellular heme and iron-sulfur cluster species, as well as mitochondrial iron and its derivatives. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

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Homology modeling : Methods and protocols

It Provides state-of-the-art methodologies and reviews of important topics in the field of homology modeling. From homology modeling in the twilight zone and improving accuracy through sequence space analysis to approaches to construct multi-protein complex models, the book explores a wide variety of uses and applications of this valuable technique.

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Histones : Methods and protocols

Provides new and updated chapters on histones for molecular biologists, biochemists and geneticists. Chapters cover protein complexes that modify chromatin either by adding or removing post-translational modifications, or by exchanging histone variants within the nucleosome. Written in the highly successful Methods in Molecular Biology series format, the chapters include brief introductions to the material, lists of necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and a Notes section which highlights tips on troubleshooting and avoiding known pitfalls.

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Heterologous protein production in CHO cells : Methods and protocols

Explores the use of Chinese hamster ovary (CHO) cells in the production of therapeutic protein products. Beyond updates on earlier methodologies, the book also delves into the genetic manipulation of CHO cells for recombinant protein production, analysis of CHO cells using proteomic and metabolomic approaches, as well as methods for the characterization of recombinant protein products, such as glycosylation and host cell protein analysis.

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Golden gate cloning : Methods and protocols

Provides an overview of the latest protocols that cover the full range of in silico, in vitro, and in vivo approaches to help researchers plan, design, and perform Golden Gate cloning experiments. The chapters in this book cover topics such as Golden Gate cloning of standardized parts; modular DNA construct design for high-throughput Golden Gate Assembly; Golden Gate cloning of Synthetic CRISPR RNA spacer sequences; Golden Gate cloning in Actinobacteria; and YeastFab cloning of toxic genes and protein expression optimization in yeast.

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Foodborne bacterial pathogens : Methods and protocols

This updated volume presents a compilation of various representative techniques and approaches currently used to study bacterial foodborne pathogens. Chapters guide the reader through bacterial pathogen detection and quantification in food, molecular, phenotypic, metabolic characterization of food pathogens, and ecology of foodborne bacterial pathogens. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.

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Food allergens : methods and protocols

Provides a comprehensive collection of methods and protocols in food allergy and food allergens studies. The selected protocols explore the study of food allergens, from recombinant production, purification procedures, IgE and T cell epitopes characterization, to allergen structure description, cellular responses, and tolerance induction, through a variety of techniques and animal models.

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Epigenome editing : Methods and protocols

This second volume details new and updated methods covering applications of epigenome editing. Chapters guide readers through general and topical reviews, DNA-binding devices, optimization of the effector domains, readout of epigenome marks, and approaches for delivery at the cellular and organismal level. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.

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Endophytic microbes: Isolation, identification, and bioactive potentials

Provides basic insight and protocols relating to endophytic microbes. Chapter are divided into five major sections detailing basic isolation, bioactive metabolites production. endophytism, isolation and identification of endophytes, bioactive potentials, and screening of metabolites.

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DNA damage detection : Methods and protocols

Explores techniques in DNA damage analysis, important not only for understanding cellular health after exposure to genotoxic agents but also for assessing cellular ability of genome maintenance. The book explores traditional DNA damage measurement methods, as well as modern fluorescence-based analysis and high throughput automatic analysis methods. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

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DNA and RNA origami : methods and protocols

Detials diverse methodological approaches on the assembly and applications of DNA origami assemblies. Chapters guide readers through different synthetic and computational methods, isolation and structural characterization of 2D and 3D DNA origami nanoarchitectures, nanophotonics, drug delivery, biophysics, and synthetic biology.Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.

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Dictyostelium discoideum : Methods and protocols

Discusses the latest advancements and techniques used to study Dictyostelium discoideum. The chapters in this book cover topics such as cytokinesis, membrane function and damage, and nutrient/bacterial processing; autophagy activation and function; protein localization, interaction, and activation; cell migration and contact guidance for quantification of cytoskeletal dynamics; and global mutational screening, transcriptome analyses, and proteomics that apply to all stages of the Dictyostelium life cycle and that may generalize to human disease models. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

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